MRI: Development of high-throughput light sheet fluorescence microscopy

俄勒冈大学（University of Oregon）开发了一种能够对大量活的多细胞标本进行三维成像的仪器，特别是作为许多生物过程研究模型的斑马鱼的胚胎和幼虫。观察细胞在早期发育、伤口愈合、疾病进展和许多其他事件中的相互作用，对我们理解正常和异常的生物功能非常有益。然而，我们缺乏将最近发展的先进三维成像技术应用于大量标本的方法，这既限制了实验的速度，也限制了结果的统计稳健性。该项目通过开发新仪器解决了这一挑战。实施这项研究将为研究生和本科生提供有价值的跨学科培训，因为它促进了光学、设备控制、计算数据分析、微生物学和发育生物学方面技能的发展。除了研究之外，该仪器还将用于与夏季本科生研究实习有关的活动，以及PI多年来共同组织的一个让社会经济上处于不利地位的高中生接触科学的日间夏令营。更广泛地说，该项目开发的仪器将提供以前未见过的活体动物细胞、组织和器官的视图，可以支持对健康和疾病的新观点。近年来，先进的三维成像方法提供了深入了解细胞的空间和时间安排。一种被称为光片荧光显微镜的高分辨率三维显微镜方法已被证明在许多小时或几天的持续时间内对活标本进行成像特别强大。然而，这种方法和类似的方法受到了吞吐量非常有限的限制，在实验运行期间只能对一个或最多几个标本进行成像。越来越明显的是，发育生物体的高度可变性，以及控制它们的机制的微妙性，要求研究能够将高精度成像应用于更多数量的标本。这激发了我们将设计和构建的仪器，它集成了高通量标本处理和实时计算图像分析与光片荧光显微镜。我们将设计的仪器能够成像和分析的特定成像目标包括肠道微生物群、免疫系统和神经元群。除此之外，还有一个大的系统列表，其中长时间的三维成像具有高统计能力将是有用的，这意味着对我们的仪器有相当大的影响。

An award is made to the University of Oregon to develop an instrument capable of three-dimensional imaging of large numbers of living multicellular specimens, especially embryos and larvae of zebrafish that serve as models for the study of many biological processes. Observation of cellular interactions during early development, wound healing, disease progression, and many other events has been enormously beneficial to our understanding of both normal and aberrant biological function. We have lacked methods, however, to apply recently developed advanced three-dimensional imaging techniques to large numbers of specimens, limiting both the rate of experiments and the statistical robustness of results. This project addresses this challenge through the development of new instrumentation. Implementing this research will provide valuable cross-disciplinary training for graduate students and undergraduates, as it fosters the development of skills in optics, device control, computational data analysis, microbiology, and developmental biology. In addition to research, the instrument will be used in activities related to summer undergraduate research internships and a day camp that the PI has co-organized for several years that exposes socioeconomically disadvantaged high school students to science. More broadly, the instrumentation developed by this project will provide previously unseen views of cells, tissues, and organs in living animals that can underpin new perspectives on health and disease.In recent years, advanced three-dimensional imaging methods have provided deep insights into the spatial and temporal arrangements of cells. A high-resolution, three-dimensional microscopy method known as light sheet fluorescence microscopy has proven itself to be particularly powerful for imaging living specimens over many-hour or several-day durations. This and similar methods, however, have suffered the limitation of very limited throughput, capable of imaging one or at most a few specimens during an experimental run. It is increasingly apparent that the high variability of developing organisms, and the subtleties of mechanisms governing them, call for studies that can apply high-precision imaging to much larger numbers of specimens. This motivates the instrument we will design and build, which integrates high-throughput specimen handling and real-time computational image analysis with light sheet fluorescence microscopy. Specific imaging targets that we will design our instrument to be capable of imaging and analyzing include the gut microbiota, immune system, and groups of neurons. Beyond this, there is a large list of systems for which long-duration three-dimensional imaging with high statistical power would be useful, implying a sizeable impact for our instrumentation.